Processing and maturation of carboxypeptidase y and alkaline phosphatase in Schizosaccharomyces pombe

Hiroyuki Mukaiyama, Tomoko Iwaki, Alimjan Idiris, Kaoru Takegawa

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Schizosaccharomyces pombe carboxypeptidase Y (CPY) is synthesized as a zymogen and transported into the vacuole where maturation and activation occurs. The 110-kDa S. pombe CPY precursor is processed twice and finally converted to a mature form consisting of polypeptides of approximately 19 and 32 kDa linked by a single disulfide bond. In Saccharomyces cerevisiae, maturation of CPY occurs mostly through the activity of vacuolar aspartyl protease Pep4p, whereas a Pep4p homolog has not been found in the S. pombe genome database. Based on analysis of protease-deficient mutants, we found that S. pombe CPY was not able to be processed or activated in isp6Δpsp3Δ double disruptants. Both Isp6p and Psp3p are subtilase-type serine proteases with related sequences. Moreover, alkaline phosphatase of S. pombe was found to be localized at the vacuolar membrane and was also unprocessed in isp6Δpsp3Δ double disruptants. Vacuolar localization of GFP-fused Isp6p and Psp3p was determined by fluorescence microscopy. These results suggest that the two serine proteases Isp6p and Psp3p are functional in the vacuole and are involved in proteolytic processing of vacuolar proteins.

Original languageEnglish
Pages (from-to)203-213
Number of pages11
JournalApplied Microbiology and Biotechnology
Volume90
Issue number1
DOIs
Publication statusPublished - Apr 1 2011

Fingerprint

Carboxypeptidases
Schizosaccharomyces
Cathepsin A
Alkaline Phosphatase
Serine Proteases
Vacuoles
Aspartic Acid Proteases
Enzyme Precursors
Fluorescence Microscopy
Disulfides
Saccharomyces cerevisiae
Peptide Hydrolases
Genome
Databases
Peptides
Membranes
Proteins

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Processing and maturation of carboxypeptidase y and alkaline phosphatase in Schizosaccharomyces pombe. / Mukaiyama, Hiroyuki; Iwaki, Tomoko; Idiris, Alimjan; Takegawa, Kaoru.

In: Applied Microbiology and Biotechnology, Vol. 90, No. 1, 01.04.2011, p. 203-213.

Research output: Contribution to journalArticle

@article{780247d3a24847a2b98851d4aa04d4e0,
title = "Processing and maturation of carboxypeptidase y and alkaline phosphatase in Schizosaccharomyces pombe",
abstract = "Schizosaccharomyces pombe carboxypeptidase Y (CPY) is synthesized as a zymogen and transported into the vacuole where maturation and activation occurs. The 110-kDa S. pombe CPY precursor is processed twice and finally converted to a mature form consisting of polypeptides of approximately 19 and 32 kDa linked by a single disulfide bond. In Saccharomyces cerevisiae, maturation of CPY occurs mostly through the activity of vacuolar aspartyl protease Pep4p, whereas a Pep4p homolog has not been found in the S. pombe genome database. Based on analysis of protease-deficient mutants, we found that S. pombe CPY was not able to be processed or activated in isp6Δpsp3Δ double disruptants. Both Isp6p and Psp3p are subtilase-type serine proteases with related sequences. Moreover, alkaline phosphatase of S. pombe was found to be localized at the vacuolar membrane and was also unprocessed in isp6Δpsp3Δ double disruptants. Vacuolar localization of GFP-fused Isp6p and Psp3p was determined by fluorescence microscopy. These results suggest that the two serine proteases Isp6p and Psp3p are functional in the vacuole and are involved in proteolytic processing of vacuolar proteins.",
author = "Hiroyuki Mukaiyama and Tomoko Iwaki and Alimjan Idiris and Kaoru Takegawa",
year = "2011",
month = "4",
day = "1",
doi = "10.1007/s00253-010-3031-3",
language = "English",
volume = "90",
pages = "203--213",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "1",

}

TY - JOUR

T1 - Processing and maturation of carboxypeptidase y and alkaline phosphatase in Schizosaccharomyces pombe

AU - Mukaiyama, Hiroyuki

AU - Iwaki, Tomoko

AU - Idiris, Alimjan

AU - Takegawa, Kaoru

PY - 2011/4/1

Y1 - 2011/4/1

N2 - Schizosaccharomyces pombe carboxypeptidase Y (CPY) is synthesized as a zymogen and transported into the vacuole where maturation and activation occurs. The 110-kDa S. pombe CPY precursor is processed twice and finally converted to a mature form consisting of polypeptides of approximately 19 and 32 kDa linked by a single disulfide bond. In Saccharomyces cerevisiae, maturation of CPY occurs mostly through the activity of vacuolar aspartyl protease Pep4p, whereas a Pep4p homolog has not been found in the S. pombe genome database. Based on analysis of protease-deficient mutants, we found that S. pombe CPY was not able to be processed or activated in isp6Δpsp3Δ double disruptants. Both Isp6p and Psp3p are subtilase-type serine proteases with related sequences. Moreover, alkaline phosphatase of S. pombe was found to be localized at the vacuolar membrane and was also unprocessed in isp6Δpsp3Δ double disruptants. Vacuolar localization of GFP-fused Isp6p and Psp3p was determined by fluorescence microscopy. These results suggest that the two serine proteases Isp6p and Psp3p are functional in the vacuole and are involved in proteolytic processing of vacuolar proteins.

AB - Schizosaccharomyces pombe carboxypeptidase Y (CPY) is synthesized as a zymogen and transported into the vacuole where maturation and activation occurs. The 110-kDa S. pombe CPY precursor is processed twice and finally converted to a mature form consisting of polypeptides of approximately 19 and 32 kDa linked by a single disulfide bond. In Saccharomyces cerevisiae, maturation of CPY occurs mostly through the activity of vacuolar aspartyl protease Pep4p, whereas a Pep4p homolog has not been found in the S. pombe genome database. Based on analysis of protease-deficient mutants, we found that S. pombe CPY was not able to be processed or activated in isp6Δpsp3Δ double disruptants. Both Isp6p and Psp3p are subtilase-type serine proteases with related sequences. Moreover, alkaline phosphatase of S. pombe was found to be localized at the vacuolar membrane and was also unprocessed in isp6Δpsp3Δ double disruptants. Vacuolar localization of GFP-fused Isp6p and Psp3p was determined by fluorescence microscopy. These results suggest that the two serine proteases Isp6p and Psp3p are functional in the vacuole and are involved in proteolytic processing of vacuolar proteins.

UR - http://www.scopus.com/inward/record.url?scp=79954897827&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79954897827&partnerID=8YFLogxK

U2 - 10.1007/s00253-010-3031-3

DO - 10.1007/s00253-010-3031-3

M3 - Article

C2 - 21153812

AN - SCOPUS:79954897827

VL - 90

SP - 203

EP - 213

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 1

ER -